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Biodiversité et adaptation au pathogène racinaire Verticillium alfalfae chez Medicago truncatula. Importance de la micro-évolution.

Mélanie Mazurier 1
1 ECI - Ecotoxicologie & Santé des écosystèmes - ECSECO
ECOLAB - Laboratoire Ecologie Fonctionnelle et Environnement, INRA - Institut National de la Recherche Agronomique
Abstract : Pathogens, animals and weeds are responsible for losses ranging from 20% to 40% of global agricultural yield. Legumes are the second most grown crops after cereals as human and animal food, and their yield is also affected by pathogens. Verticillium alfalfae is a soil-borne pathogen causing high yield losses in alfalfa fields (Medicago sativa), the most worldwide legume forage crop grown. To date, there is no chemical control and crop breeding is the best way to protect alfalfa against V. alfalfae. However, Medicago sativa has a complex genome (allogamous and tetraploid), therefore in this work the model legume Medicago truncatula (diploid and self-fertile) was used to investigate genetic mechanisms involved in V. alfalfae V31-2 (Va V31-2) resistance. First, several sources of resistance were identified in M. truncatula biodiversity by assessing disease symptoms development in 261 lines from the Mediterranean basin inoculated by V. alfalfae V31-2. Reisolation of V. alfalfae V31-2 in M. truncatula stems was also led. These analyses revealed a great biodiversity of M. truncatula response towards Va V31-2. A genome wide association study (GWAS) on various disease phenotypes pinpointed several quantitative trait loci (QTL): one of them colocalized with a previous major QTL on chromosome 7 detected on LR4 and LR5 RILs populations. Both phenotypic and genetic analyses thus suggest the occurrence of different resistance mechanisms in M. truncatula populations towards V. alfalfae. Resistant candidate genes towards Va V31-2 were also identified by GWAS using 90 accessions of M. truncatula Soliman Tunisian population. These candidate genes are different from the pinpointed candidate gene of our previous GWAS analysis. These results might underline a local adaptation towards Verticillium. Consistencies between GWAS results with previous QTL and transcriptomic data led us to perform the functional validation with the candidate genes localized on chromosome 7. An original in vitro inoculation system of M. truncatula transgenic roots was used to validate Medtr7g070480 (gene encoding a SEC14 protein) as a key player towards V. alfalfae V31.2 in M. truncatula. Silencing the SEC14 gene using artificial microRNA in A17 (resistant) and F83005.5 (susceptible) lines decreases Va V31-2 colonization rate on transgenic roots whereas overexpressing MtSEC14 increases the colonization rate in A17. This MtSEC14 gene appears as a disease susceptibility gene. Moreover, 18.5% of the disease variation in the studied M. truncatula accessions is explained by the unique non-synonymous polymorphism between A17 and F83005.5 in MtSEC14. Root expression patterns of the SEC14 gene one day after inoculation by Va V31-2 was also analyzed in response to inoculation among a panel of 14 susceptible and resistant M. truncatula accessions of diverse geographic origins. No significant differences were found. At the same time, orthologous resistance candidate genes towards Va V31.2 in several other model species (Lotus japonicus, Arabidopsis thaliana, Nicotiana sp.) were discovered. Pathogenicity of Va V31-2 in these species was monitored to use them as potential model for functional validation. Only one orthologous gene of MtSEC14 has been identified in Medicago sativa. Because of a great synteny between M. truncatula and M. sativa, our work could be useful for alfalfa breeding. In this work and for the first time, a susceptibility gene involved in a quantitative resistance against microorganisms in Legumes was identified.chez les Légumineuses.
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Submitted on : Monday, November 19, 2018 - 3:52:04 PM
Last modification on : Thursday, October 15, 2020 - 4:07:27 AM


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Mélanie Mazurier. Biodiversité et adaptation au pathogène racinaire Verticillium alfalfae chez Medicago truncatula. Importance de la micro-évolution.. Ecologie, Environnement. Université Paul Sabatier (Toulouse 3), 2018. Français. ⟨tel-01927066⟩



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